Clinical And Mechanism Study On Fuzheng Kangai Decoction In The Treatment Of Ovarian Cance

Purpose:To observe the clinical efficacy and safety of Fuzheng Kangai Decoction combined with TC regimen in the treatment of ovarian cancer after operation through clinical trials;to study the possible mechanism of Fuzheng Kangai Decoction in the treatment of ovarian cancer through the method of network pharmacology;through the animal experiment,it was evaluated from the molecular level that Fuzheng Kangai Decoction combined with Cisplatin inhibitted the proliferation of ovarian cancer cells,promoted the apoptosis of ovarian cancer cells,and regulated the immune function of the body,and discussed the mechanism of Fuzheng Kangai Decoction combined with cisplatin in the treatment of ovarian cancer based on the PI3K/Akt signaling pathway.To provide reference for clinical application of Fuzheng Kangai Decoction in the treatment of ovarian cancer.Material and method:1.Paper 1: In this study,from September 2019 to November 2021,123 ovarian cancer patients after operation who met the inclusion criteria were selected from the patients with ovarian cancer who were hospitalized in the Department of Gynecology,the Cancer Hospital of Liaoning province,and they were divided into two groups by random number table method,an observation group of 62 cases,a control group of 61 cases.During the trial,3 patients were excluded because they did not take medicines or examinations as prescribed.A total of 120 cases were finally included in the trial,including 60 cases in the observation group and 60 cases in the control group.The treatment plan of the observation group was Fuzheng Kangai Decoction combined with TC chemotherapy(intravenous infusion of paclitaxel combined with carboplatin),and the treatment plan of the control group was TC chemotherapy alone(intravenous infusion of paclitaxel combined with carboplatin).Both groups of patients were routinely treated with antiemetic,stomach protection,hydration,diuresis,and allergy prevention during the process of chemotherapy.A cycle of chemotherapy was 21 days,the two groups were intervented with a total of 2 cycles.The observation indicators were: general informations such as age,height,weight,BMI,pathological type,clinical stage,etc;KPS score,Traditional Chinese Medicine syndrome score and quality of life score of patients before and after the treatment,as well as serum tumor markers,cellular immune function,blood routine,biochemical indicators,nausea and vomiting,allergies and other side effects of the chemotherapy.Comparision the efficacy and safety of the two treatment regimens.2.Paper 2:Screening and predicting targets of active ingredients in Fuzheng Kangai Decoction through TCMSP database and reviewed the literature,and screening ovarian cancer disease’s targets through Gene Cards,OMIM,Pharmgkb,TTD,and Drug Bank databases.The targets of the Fuzheng Kangai Decoction’s active ingredients and the disease’s targets of ovarian cancer were intersected to construct a protein interaction network(PPI),and GO enrichment analysis and KEGG pathway enrichment analysis were carried out on the intersection targets to explore the mechanisms of Fuzheng Kangai Decoction in the treatment of ovarian cancer.3.Paper 3: 32 SPF female BALB/c-nu nude mice were selected to construct an ovarian cancer xenograft model,and the 32 nude mice were randomly divided into 4 groups: control group,traditional Chinese medicine group,cisplatin group,and traditional Chinese medicine combined with cisplatin group,which had 8 mice in each group.The ovarian cancer tumor-bearing nude mice in the traditional Chinese medicine group were given Fuzheng Kangai Decoction by gavage each day and intraperitoneal injection of normal saline every two days,the ovarian cancer tumor-bearing nude mice in the cisplatin group were given intraperitoneal injection of cisplatin for injection every two days and normal saline by gavage each day,the ovarian cancer tumor-bearing nude mice in the traditional Chinese medicine combined with cisplatin group were given Fuzheng Kangai Decoction by gavage each day and intraperitoneal injection of cisplatin for injection every two days,and the ovarian cancer tumor-bearing nude mice in the control group were given normal saline by gavage each day and intraperitoneal injection of normal saline every two days.During the administration period,the mental state,activity and feeding situation of nude mice were observed every day,and the tumor volume was measured every other day.Nude mice were killed after 2 weeks of intervention,serum,spleen and tumor body of each group of nude mice were collected for future use,and the tumor body weight was weighed and photographed for record.In order to evaluate the inhibitory effect of Fuzheng Kangai Decoction combined with cisplatin on ovarian cancer,the tumor inhibition rate was calculated,the Ki67 level in the transplanted tumor tissue was detected by immunohistochemical method,and the apoptosis rate of the transplanted tumor tissue was detected by TUNEL method;HE staining was used to observe the the effects of each group of drugs on the tissue histopathological morphology of transplanted tumors;ELISA was used to detect the contents of TNF-α and IL-2 in serum,and the immune organ index was calculated to evaluate the effects of Fuzheng Kangai Decoction combined with cisplatin on the immune functions of ovarian cancer tumor-bearing nude mice;the method of Western blot was used to detect the expression levels of PTEN,PI3 K,p-Akt and Akt proteins in the transplanted tumor tissue to evaluate the effects of Fuzheng Kangai Decoction combined with cisplatin on PI3K/Akt signaling pathway.Results:1.Paper 1: After treatment,the KPS score,Traditional Chinese Medicine syndrome score,Traditional Chinese Medicine Syndrome single integral and quality of life score of the two groups were improved compared with those before treatment,and the differences were statistically significant between the observation group and the control group(P<0.05);After treatment,the efficacy of KPS score and the TCM syndromes in the observation group were better than those in the control group,and the differences were statistically significant(P<0.05);After treatment,the levels of tumor markers of CA125 and HE4 in the two groups were significantly decreased compared with those before treatment,and the differences were statistically significant between the observation group and the control group(P<0.05).After treatment,the cellular immune function of the two groups was significantly improved compared with that before treatment,and the difference was statistically significant between the observation group and the control group(P<0.05).In terms of reducing the toxicity and safety of chemotherapy,the incidences of leukopenia,thrombocytopenia,increased creatinine,nausea and vomiting in the observation group were better than those in the control group,and the differences were statistically significant(P<0.05);The comparisons of the incidences of neutropenia,decreased hemoglobin,increased bilirubin,increased alanine aminotransferase,increased alkaline phosphatase,diarrhea,allergy,peripheral neuropathy,there were no significant differences between the observation group and the control group(P>0.05).2.Paper 2: In total,203 main active ingredients and 503 corresponding drug targets of Fuzheng Kangai Decoction and 7502 disease targets of ovarian cancer were screened out in this study.408 intersection targets of drugs and disease were selected to establish a PPI network.GO enrichment analysis was performed to obtain 3056 biological process(BP)entries,129 cellular component(CC)entries,and 293 molecular function(MF)entries.KEGG pathway enrichment analysis was performed to obtain 186 KEGG pathways,among which the PI3K-Akt signaling pathway had the most enriched core target genes.3.Paper 3:(1)Compared with the control group,the tumor weight of the traditional Chinese medicine group,the cisplatin group,and the traditional Chinese medicine combined with cisplatin group were decreased(P<0.05),the tumor volume growth curve decreased(P<0.05),and the tumor inhibition rate increased;Compared with the traditional Chinese medicine group or the cisplatin group,the traditional Chinese medicine combined with cisplatin group had a more significant decrease in tumor weight and a more significant decrease in the growth curve(P<0.05).(2)Compared with the control group,the spleen index of nude mice in the traditional Chinese medicine group,the cisplatin group,and the traditional Chinese medicine combined with cisplatin group all increased,and the difference of spleen index in the traditional Chinese medicine group was statistically significant(P<0.05).Compared with the control group,the traditional Chinese medicine group,the cisplatin group,the traditional Chinese medicine combined with cisplatin group,the levels of TNF-α and IL-2 in serum were increased,the traditional Chinese medicine combined with cisplatin group increased more significantly(P<0.05).(3)Compared with the control group,the average optical density in the traditional Chinese medicine group,the cisplatin group,and the traditional Chinese medicine combined with cisplatin group were significantly decreased,and the apoptosis rate was significantly increased(P<0.05);Compared with the traditional Chinese medicine group or the cisplatin group,the average optical density decreased and the apoptosis rate increased significantly of the traditional Chinese medicine combined with cisplatin group(P<0.05).The results of HE staining also confirmed that the traditional Chinese medicine combined with cisplatin group had the strongest effect on promoting the apoptosis of ovarian cancer cells and inhibiting the proliferation of ovarian cancer cells.(4)Compared with the control group,the expression of PTEN protein in the traditional Chinese medicine group,cisplatin group,and traditional Chinese medicine combined with cisplatin group was significantly increased,while the protein expression of PI3 K and p-Akt was significantly decreased,and the differences were statistically significant(P<0.05);Compared with the traditional Chinese medicine group or the cisplatin group,the traditional Chinese medicine combined with cisplatin group up-regulated the expression of PTEN protein of the transplanted tumor issue,and down-regulated the protein expression of PI3 K and p-Akt more significantly(P<0.05).Conclusion:1.Paper 1:(1)The plan of Fuzheng Kangai Decoction combined with TC has significant advantages in improving the KPS score,reducing the score of Traditional Chinese Medicine syndromes and individual Traditional Chinese Medicine symptom scores,reducing the quality of life score of patients who were diagnosed with ovarian cancer after operation.(2)The plan of Fuzheng Kangai Decoction combined with TC has significant advantages in reducing the levels of tumor markers of CA125 and HE4 and regulating the body’s immunity.(3)The plan of Fuzheng Kangai Decoction combined with TC has significant advantages in reducing toxicity and side effects caused by chemotherapy and improving the safety of chemotherapy.2.Paper 2:Fuzheng Kangai Decoction may treat the ovarian cancer through multiple targets and multiple pathways,and the mechanism of Fuzheng Kangai Decoction in the treatment of ovarian cancer may be related to the regulation of PI3K/Akt signaling pathway,TNF signaling pathway,p53 signaling pathway,IL-17 signaling pathway and other signaling pathways.3.Paper 3:(1)Fuzheng Kangai Decoction combined with cisplatin can inhibit the growth of HO-8910 PM tumor-bearing nude mice’s transplanted tumor,inhibit the proliferation of ovarian cancer cells,and promote the apoptosis of ovarian cancer cells.(2)Fuzheng Kangai Decoction combined with cisplatin can enhance the immune function of the body.(3)Fuzheng Kangai Decoction combined with cisplatin in the treatment of ovarian cancer may be related to the mediating of PI3K/Akt signaling pathway by up-regulating PTEN.